Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing
RNA editing is a significant mechanism underlying genetic variation and protein molecule alteration; C-to-U RNA editing, specifically, is important in the regulation of mammalian genetic diversity. The ability to define and limit accesses of enzymatic machinery to avoid the modification of unintende...
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MDPI AG
2025-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/15/1/136 |
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| author | Sonali Bhakta Hiroko Kodama Masakazu Mimaki Toshifumi Tsukahara |
| author_facet | Sonali Bhakta Hiroko Kodama Masakazu Mimaki Toshifumi Tsukahara |
| author_sort | Sonali Bhakta |
| collection | DOAJ |
| description | RNA editing is a significant mechanism underlying genetic variation and protein molecule alteration; C-to-U RNA editing, specifically, is important in the regulation of mammalian genetic diversity. The ability to define and limit accesses of enzymatic machinery to avoid the modification of unintended targets is key to the success of RNA editing. Identification of the core component of the apoB RNA editing holoenzyme, APOBEC, and investigation into new candidate genes encoding other elements of the complex could reveal further details regarding APOBEC-mediated mRNA editing. Menkes disease is a recessive X-chromosome-linked hereditary syndrome in humans, caused by defective copper metabolism due to mutations in the <i>ATP7A</i> gene, which encodes a copper transport protein. Here, we generated plasmids encoding the MS2 system and the APOBEC1 deaminase domain and used a guide RNA with flanking MS2 sites to restore mutated <i>Atp7a</i> in fibroblasts from a macular mouse model of Menkes disease withs T>C mutation. Around 35% of the mutated C nucleotide (nt) was restored to U, demonstrating that our RNA editing system is reliable and has potential for therapeutic clinical application. RNA base editing via human RNA-guided cytidine deaminases is a potentially attractive approach for in vivo therapeutic application and provides opportunities for new developments in this field. |
| format | Article |
| id | doaj-art-577cc469b83548b2b95789d5513af0dd |
| institution | Kabale University |
| issn | 2218-273X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
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| series | Biomolecules |
| spelling | doaj-art-577cc469b83548b2b95789d5513af0dd2025-01-24T13:25:19ZengMDPI AGBiomolecules2218-273X2025-01-0115113610.3390/biom15010136Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA EditingSonali Bhakta0Hiroko Kodama1Masakazu Mimaki2Toshifumi Tsukahara3Bioscience, Biotechnology and Biomedical Engineering Research Area, Japan Advanced Institute of Science and Technology, Nomi 923-1211, JapanGeneral Medical Education and Research Center, Teikyo University School of Medicine, Tokyo 173-0003, JapanDepartment of Pediatrics, Teikyo University School of Medicine, Tokyo 173-0003, JapanBioscience, Biotechnology and Biomedical Engineering Research Area, Japan Advanced Institute of Science and Technology, Nomi 923-1211, JapanRNA editing is a significant mechanism underlying genetic variation and protein molecule alteration; C-to-U RNA editing, specifically, is important in the regulation of mammalian genetic diversity. The ability to define and limit accesses of enzymatic machinery to avoid the modification of unintended targets is key to the success of RNA editing. Identification of the core component of the apoB RNA editing holoenzyme, APOBEC, and investigation into new candidate genes encoding other elements of the complex could reveal further details regarding APOBEC-mediated mRNA editing. Menkes disease is a recessive X-chromosome-linked hereditary syndrome in humans, caused by defective copper metabolism due to mutations in the <i>ATP7A</i> gene, which encodes a copper transport protein. Here, we generated plasmids encoding the MS2 system and the APOBEC1 deaminase domain and used a guide RNA with flanking MS2 sites to restore mutated <i>Atp7a</i> in fibroblasts from a macular mouse model of Menkes disease withs T>C mutation. Around 35% of the mutated C nucleotide (nt) was restored to U, demonstrating that our RNA editing system is reliable and has potential for therapeutic clinical application. RNA base editing via human RNA-guided cytidine deaminases is a potentially attractive approach for in vivo therapeutic application and provides opportunities for new developments in this field.https://www.mdpi.com/2218-273X/15/1/136RNA editingmacular mousefibroblastAPOBEC 1MS2 systemguide RNA |
| spellingShingle | Sonali Bhakta Hiroko Kodama Masakazu Mimaki Toshifumi Tsukahara Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing Biomolecules RNA editing macular mouse fibroblast APOBEC 1 MS2 system guide RNA |
| title | Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing |
| title_full | Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing |
| title_fullStr | Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing |
| title_full_unstemmed | Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing |
| title_short | Restoration of Genetic Code in Macular Mouse Fibroblasts via APOBEC1-Mediated RNA Editing |
| title_sort | restoration of genetic code in macular mouse fibroblasts via apobec1 mediated rna editing |
| topic | RNA editing macular mouse fibroblast APOBEC 1 MS2 system guide RNA |
| url | https://www.mdpi.com/2218-273X/15/1/136 |
| work_keys_str_mv | AT sonalibhakta restorationofgeneticcodeinmacularmousefibroblastsviaapobec1mediatedrnaediting AT hirokokodama restorationofgeneticcodeinmacularmousefibroblastsviaapobec1mediatedrnaediting AT masakazumimaki restorationofgeneticcodeinmacularmousefibroblastsviaapobec1mediatedrnaediting AT toshifumitsukahara restorationofgeneticcodeinmacularmousefibroblastsviaapobec1mediatedrnaediting |